Mycoplasma Detection Kit (Probe qPCR) servicebio G1903-50
$50.00
In stock
SKU
G1903-50
Product Information
| Product Name: Mycoplasma Detection Kit (Probe qPCR) | Cat. No .: G 1903-50 |
| Spec.: 50 rxns | G1903-100 |
| 100 rxns |
Product Description
This product is based on Taqman probe method to detect 8 common mycoplasma contaminations in cell culture and other biological materials, which is characterized by simple operation, high sensitivity and high specificity.Detectable samples are supernatants from cell cultures performed 3-6 days after inoculation; If the samples are cell suspensions or in the presence of qPCR inhibitors, DNA needs to be extracted before the reaction is performed. This kit also provides PositiveControl, which can be added to the sample before the qPCR reaction to prevent false negative results if you are unsure whether the sample contains a qPCR inhibitor.
Storage and Shipping Conditions
Ship with wet ice; Store at -20℃ away from light for 12 months.
Product Content
| Component Number: G1903-1 | Component: 2× Myco Universal Blue qPCR Master Mix |
| G1903-50: 500 µL | G1903-100: 1 mL |
| Component Number: G1903-2 | Component: Myco Primer s & Probe Mix |
| G1903-50: 60 µL | G1903-100: 120 µL |
| Component Number: G1903-3 | Component: Positive Control |
| G1903-50: 50 µL | G1903-100: 100 µL |
| Manual | 1 pc |
Pre-experiment preparation
1. Real Time PCR Amplifier.
2. qPCR tube or plate for experiments.
Assay Protocol
1. Sample Preparation:
Adherent cells: If commonly used cell culture medium is used, 1 µL of medium supernatant can be taken and added directly to the qPCR reaction system when cells are inoculated or changed for more than 3 days.
Suspension cells: DNA needs to be extracted, then qPCR reaction.
Reaction Inhibitors: If qPCR inhibitors are present in the cell culture solution, DNA needs to be extracted before performing the qPCR reaction.
2. Suggested qPCR reaction system:
| Component: 2× Myco Universal Blue qPCR Master Mix a | 20 μL rxn: 10 μL |
| Component: Myco Primer & Probe Mix | 20 μL rxn: 1.2 μL |
| Component: Sampla b | 20 μL rxn: 1 μL |
| Component: Nuclease-Free Water | 20 μL rxn: Add to 20 μL |
a. The ROX Reference Dye is not included in the qPCR Master Mix, when setting up the qPCR program, select the ROX channel as None; if you need to add the ROX Reference Dye, please purchase the 50× ROX Reference Dye and use it at a final concentration of 1×.
b. If you are not sure whether the sample contains qPCR inhibitors, add 0.5 µL of Positive Control to the qPCR reaction system at the same time as adding the sample; If Negative Control is required, the sample can be replaced with Nuclease-Free Water or without any template.
3. qPCR reaction program (can be adjusted appropriately according to the model):
A. Two-step method
B. Three-step method
| Stage: Stage 1 | Step: premutability |
| C ycle: 1 | T emperature: 95℃ |
| Time: 30 sec | Stage: Stage 2 |
| Step: denaturation | C ycle: 40 |
| T emperature: 95℃ | Time: 15 sec |
| Annealing/Extension | 60℃ |
| 30 sec a | Stage: Stage |
| Step: Step | C ycle: C ycle |
| T emperature: T emperature | Time: Time |
| Stage: Stage 1 | Step: premutability |
| C ycle: 1 | T emperature: 95℃ |
| Time: 30 sec | Stage: Stage 2 |
| Step: denaturation | C ycle: 40 |
| T emperature: 95℃ | Time: 15 sec |
| Annealing | 55-65℃ |
| 10 sec | Extension |
| 72℃ | 30 sec a |
To improve amplification specificity, use a two-step program or increase the annealing temperature;
To improve amplification efficiency, use a three-step program or extend the extension time.
4. qPCR reaction parameters:
The reporter fluorescent moiety is FAM and the quenching fluorescent moiety is None selected.
Note
1. After thawing the reagent, mix it upside down gently before use. Do not swirl and mix it to avoid bubbles.
2. When preparing the reaction solution, keep the reagents on ice.
3. Please use new disposable tips for preparation and dispensing of reaction solution to avoid contamination between samples as much as possible.
4. Avoid repeated freezing and thawing and try to use it within one month after thawing.
5. For your health and safety, please wear lab coat and gloves during operation.
Compatible Models
ABI: 5700, 7000, 7300, 7700, 7900, 7900HT, 7900 HT Fast, StepOne™, StepOne Plus™, 7500/7500 Fast, ViiA 7™, QuantStudio™, PikoRealTM Cycler;
Stratagene: Mx3000P®, 3005P™, 4000™;
Bio-Rad: CFX96™, CFX384™, iCycler iQ™, iQ5™, MyiQ™, MiniOpticon™, Opticon®, Opticon 2, Chromo4™;
Eppendorf: Realplex 2s, Mastercycler® ep, realplex;
IIIumina: Eco QPCR
Cepheid: SmartCycler®
Qiagen Corbett: Rotor-Gene®
Roche: LightCycler™
Takara: Thermal Cycler Dice
Analytikjena: qTOWER
qTOWER: LineGene
For Research Use Only!
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